Cationic cardiolipin analoges and its use thereof

Abstract

The invention provides cationic cardiolipin compounds, and methods for synthesizing and using them in liposomal formulation, gene transfection, etc. In particular, the invention provides liposomes comprising cationic cardiolipin analog, pharmaceutical compositions comprising cationic cardiolipin analogs, and methods of using such liposomes and compositions, in delivering active pharmaceutical agents to treat human and animal diseases and/or in diagnostic assays.

1 . A cationic cardiolipin analog having the general structure I.  
       
         
           
           
               
               
           
         
       
       wherein Z 1  and Z 2  are the same or different and are —O—C(O)—, —O—, —S—, or —NH—C(O)—; 
 R 1 , R 2 , R 3 , and R 4  are the same or different and are H, C 1  to C 32  saturated or unsaturated alkyl, alkenyl, or alkynyl, groups, optionally hydroxylated, aminenated, thiolated, epoxylated, cyclolated, PEGylated, halogenated, or substituted with combinations thereof;  
 R 5  and R 6  are the same or different and can be either absent or comprise a linker comprising a C 1 -C 32  alkyl, substituted alkyl, cycloalkyl, substituted cycloalky, or an alkyloxy or substituted alkyloxy group such as a PEGylated ether containing from 1 to 500 PEG (polyethylene glycol) units;  
 R 7  is hydrogen, alkyl, substituted alkyl, alkyloxy, substituted alkyloxy, cycloalkyl, substituted cycloalkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, optionally hydroxylated, aminenated, thiolated, epoxylated, cyclolated, PEGylated, halogenated, or substituted with combinations thereof; or an alkyloxy or substituted alkyloxy group such as PEGylated ether containing from 1 to 500 PEG (polyethylene glycol) units;  
 an amino acid, folic acid, a peptide, a peptidomimetic moiety, a dipeptide, a polypeptide, a protein, a carbohydrate, a saccharide or polysaccharide, a polyamine, a heterocyclic, a nucleoside, or a polynucleotide;  
 the R 8  groups are the same or different and are C 1  to C 25  saturated or unsaturated alkyl, alkyloxy, substituted alkyl, or substituted alkyloxy;  
 X is a non-toxic anion.  
 
     
     
         2 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ether having the structure of formula II:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 32  alkyl, alkenyl, or alkynyl.  
     
     
         3 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ester having the structure of formula III:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 31 , alkyl, alkenyl, or alkynyl.  
     
     
         4 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ether having the structure of formula IV:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 32  alkyl, alkenyl, or alkynyl.  
     
     
         5 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ester having the structure of formula V:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 31  alkyl, alkenyl, or alkynyl.  
     
     
         6 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ether having the structure of formula VI:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 32  alkyl, alkenyl, or alkynyl.  
     
     
         7 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ester having the structure of formula VII:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 31  alkyl, alkenyl, or alkynyl.  
     
     
         8 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ether having the structure of formula VIII:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 32  alkyl, alkenyl, or alkynyl.  
     
     
         9 . The analog of  claim 1 , wherein the analog is a cationic cardiolipin ester having the structure of formula IX:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 31 , alkyl, alkenyl, or alkynyl.  
     
     
         10 . A cationic cardiolipin analog having the general structure X.  
       
         
           
           
               
               
           
         
       
       wherein Z 1  and Z 2  are the same or different and are —O—C(O)—, —O—, —S—, or —NH—C(O)—; 
 R 1 , R 2 , R 3 , and R 4  are the same or different and are H, C 1  to C 32  saturated or unsaturated alkyl, alkenyl, or alkynyl, groups, optionally hydroxylated, aminenated, thiolated, epoxylated, cyclolated, PEGylated, halogenated, or substituted with combinations thereof;  
 R 5  and R 6  are the same or different and can be either absent or comprise a linker comprising a C 1 -C 32  alkyl, substituted alkyl, cycloalkyl, substituted cycloalky, or an alkyloxy or substituted alkyloxy group such as PEGylated ether containing from 1 to 500 PEG (polyethylene glycol) units;  
 the R 7  groups can be the same or different and are hydrogen, alkyl, substituted alkyl, alkyloxy, substituted alkyloxy, cycloalkyl, substituted cycloalkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, optionally hydroxylated, aminenated, thiolated, epoxylated, cyclolated, PEGylated, halogenated, or substituted with combinations thereof; or an alkyloxy or substituted alkyloxy group such as PEGylated ether containing from 1 to 500 PEG (polyethylene glycol) units; an amino acid, a peptide, a peptidomimetic moiety, a dipeptide, a polypeptide, a protein, a carbohydrate, a saccharide or polysaccharide, a polyamine, a heterocyclic, a nucleoside, folic acid or a polynucleotide;  
 the R 8  groups are the same or different and are C 1  to C 25  saturated or unsaturated alkyl, alkyloxy, substituted alkyl, or substituted alkyloxy;  
 X is a non-toxic anion.  
 
     
     
         11 . The analog of  claim 10 , wherein the analog is a cationic cardiolipin ether having the structure of formula XI:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 32  alkyl, alkenyl, alkynyl.  
     
     
         12 . The analog of  claim 10 , wherein the analog is a cationic cardiolipin ester having the structure of formula XII:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 31  alkyl, alkenyl, or alkynyl.  
     
     
         13 . A cationic cardiolipin analog having the general structure XIII.  
       
         
           
           
               
               
           
         
       
       wherein Z 1  and Z 2  are the same or different and are —O—C(O)—, —O—, —S—, or —NH—C(O)—; 
 R 1 , R 2 , R 3 , and R 4  are the same or different and are H, C 1  to C 32  saturated or unsaturated alkyl, alkenyl, or alkynyl, groups, optionally hydroxylated, aminenated, thiolated, epoxylated, cyclolated, PEGylated, halogenated, or substituted with combinations thereof;  
 R 5  and R 6  are the same or different and can be either absent or comprise a linker comprising a C 1 -C 32  alkyl, substituted alkyl, cycloalkyl, substituted cycloalky, or an alkyloxy or substituted alkyloxy group such as PEGylated ether containing from 1 to 500 PEG (polyethylene glycol) units;  
 R 7 , R 9 , R 10  and R 11 , are the same or different and are hydrogen, alkyl, substituted alkyl, alkyloxy, substituted alkyloxy, cycloalkyl, substituted cycloalkyl, alkenyl, alkynyl, alkanoyl, alkenoyl, alkynoyl, optionally hydroxylated, aminenated, thiolated, epoxylated, cyclolated, PEGylated, halogenated, or substituted with combinations thereof; or an alkyloxy or substituted alkyloxy group such as PEGylated ether containing from 1 to 500 PEG (polyethylene glycol) units; an amino acid, a peptide, a peptidomimetic moiety, a dipeptide, a polypeptide, folic acid, a protein, a carbohydrate, a saccharide or polysaccharide, a polyamine, a heterocyclic, a nucleoside, or a polynucleotide;  
 R 8  is C 2 -C 32  alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkyloxy or substituted alkyloxy;  
 X is a non-toxic anion.  
 
     
     
         14 . The analog of  claim 13  having the structure of formula XIV:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, C 1  to C 32  alkyl, alkenyl, or alkynyl.  
     
     
         15 . The analog of  claim 13  having the structure of formula XV:  
       
         
           
           
               
               
           
         
       
       wherein R 1 , R 2 , R 3 , and R 4  are the same or different and are H, or C 1  to C 31  alkyl, alkenyl, or alkynyl.  
     
     
         16 . A lipid composition, comprising a cationic cardiolipin analog according to any of claims  1 ,  10  or  13 .  
     
     
         17 . The composition of  claim 16 , further comprising one or more co-lipids.  
     
     
         18 . The composition of claims  17 , wherein the co-lipids comprise neutral lipids.  
     
     
         19 . The composition of  claim 18 , wherein the molar ratio of the cationic cardiolipin analog to neutral co-lipids is between about 9:1 to about 1:9.  
     
     
         20 . The composition of  claim 17 , wherein the co-lipids are selected from a group consisting of phosphatidylethanolamine, phosphatidylcholine, sphingomyelin, sterol, tocopherol and cationic lipids.  
     
     
         21 . The composition of  claim 20 , wherein the cationic lipid is selected from a group consisting of N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium choloride (DOTMA) and 1,2,-dioleoyloxy-3-(trimethylammonium)-propane (DOTAP).  
     
     
         22 . The composition of  claim 20 , wherein the phosphatidylcholine is selected from a group consisting of dimyristoylphosphatidyl choline, distearoylphosphatidylcholine, dioleylphosphatidyl choline, dipalmitoylphosphatidylcholine, diarachidonoylphosphatidylcholine, egg phosphatidylcholine, soy phosphatidylcholine, hydrogenated soy phosphatidylcholine, and mixtures thereof.  
     
     
         23 . The composition of  claim 20 , wherein the sterol is selected from a group consisting of cholesterol, derivatives of cholesterol, coprostanol, cholestanol, cholestane, cholesterol hemisuccinate, cholesterol sulfate, and mixtures thereof.  
     
     
         24 . The composition of  claim 16 , wherein the composition comprises liposomes.  
     
     
         25 . The composition of  claim 24 , wherein the liposomes have a net neutral, negative or positive charge.  
     
     
         26 . The composition of  claim 24 , wherein the liposomes are coated with biodegradeable polymers.  
     
     
         27 . The composition of  claim 24 , wherein the liposomes comprise unilamellar vesicles, multilamellar vesicles or mixtures thereof.  
     
     
         28 . The composition of  claim 24 , wherein the liposomes have a diameter of about 1 micron to 500 nm.  
     
     
         29 . The composition of  claim 16 , further comprising a physiological acceptable vehicle.  
     
     
         30 . The composition of  claim 16 , further comprising one or more active agents.  
     
     
         31 . The composition of  claim 30 , wherein the active agent is selected from a group consisting of nucleoside analogue, nucleotide analogue, corticosteroid, non-steroidal anti-inflammatory agent, bioactive lipid, anticancer drugs, antibiotic, antifingal agent, oxidant, antiviral agent, nucleoside protein, peptide, polypeptide, polynucleotide, antisense polynucleotide, oligonucleotide and antisense oligonucleotide.  
     
     
         32 . The composition of  claim 16 , further comprising a targeting agent.  
     
     
         33 . The composition of  claim 32 , wherein the targeting agent is selected from a group consisting of a carbohydrate, protein and antibody.  
     
     
         34 . A method of treating a disease, comprising administering the composition of  claim 16  to a patient in need of treatment in an amount and at a location sufficient to treat the disease within the patient.  
     
     
         35 . The method of  claim 34 , wherein the disease is cancer or a viral infection.  
     
     
         36 . A method of introducing an active agent into a cell, comprising (a) preparing a composition comprising a cationic cardiolipin of  claim 16  and said active agent (b) contacting said cell with said composition whereby said active agent is taken up into said cell.  
     
     
         37 . The method of  claim 36 , wherein said contacting step occurs in vitro.  
     
     
         38 . The method of  claim 36 , wherein said contacting step occurs in vivo.  
     
     
         39 . A method of transfecting a cell with a polynucleotide, comprising (a) preparing a composition comprising a cationic cardiolipin analog of  claim 16  and said polynucleotide (b) contacting said cell with said composition whereby said polynucleotide is taken up into said cell.  
     
     
         40 . The method of  claim 39 , wherein the cells are in vitro.  
     
     
         41 . The method of  claim 39 , wherein the cells are in vivo.  
     
     
         42 . A kit for transfecting cells, said kit comprising a cationic cardiolipin of  claim 16  and one or more elements selected from the group consisting of a polynucleotide, instructions for formulating the polynucleotide and the cationic cardiolipin into a preparation, instructions for transfecting cells using the cationic cardiolipin, reagents for facilitating transfection, containers for storing the cationic cardiolipin, containers for storing the polynucleotide, containers for storing the reagents, containers for storing a preparation including the cationic cardiolipin and polynucleotide, or containers for preparing the preparation, and materials to facilitate transfection.